FUNDAMENTALS OF STERILE FILTRATION Selection and Implementation

Millipore Corporation Randy Wilkins Technical Consulting Manager

Agenda
Selection criteria and techniques
Vmax test
Principles Practical application Benefits and limitations

Filter system sterilization

Considerations for successful SIP

Integrity testing
Current regulatory perspectives Practical solutions

Four Step Sterile Filter Selection

1. Materials Compatibility
Chemical compatibility Sterilization compatibility Adsorption considerations

2. Retention
Sterilizing-grade Confirmed with specific process fluid and operating parameters
Generally 0.2 or 0.1 um rated

3. Flow Rate
Can be estimated from vendor flow/dP data Understanding how flow/dP changes during processing

4. Capacity

Capacity Plugging Mechanisms

Cake formation Complete pore plugging Gradual pore plugging

Factors Controlling Plugging

High Particle to Pore Size Ratio 1

Cake Filtration Complete Pore Blocking Gradual Pore Plugging

Low

Low

Concentration

High

The filtration mechanism depends on: Particle to Pore Size Ratio Particle Concentration

Plugging Models
For identical particles & uniform pores
Different models depending on where particles are deposited

Monolayer Adsorption

Gradual Pore Plugging

Complete Pore Blocking

Caking Filtration

<<1

<1 ~1 Particle/Pore size Ratio

>1

For particle & pore size distributions

One mechanism may dominate hydraulics-apply simple model Mechanism may change over filtration-apply simple model to section of data Mixed mechanism hydraulics-apply more complex plugging models

Plugging Model Behavior

Resistance or permeability changes
Adsorption - no change Gradual - delayed change Complete - slight delay, then rapid change Cake - steady change
Plugging Models

10.0 9.0 8.0 7.0 6.0 5.0 4.0 3.0 2.0 1.0 0 200 400 L/m2 600

R/Ro

Adsorption Gradual Complete Cake

800

1000

Plugging Models
Mathematical Models Cake Formation
t/V = CV + D

Cake Formation
Particles accumulate on filter surface Hard, non deformable particles

Complete Pore Blocking

Particles completely block pore Soft deformable particles

Complete Pore Blocking

V=E(1 - e-kt)

Gradual Pore Blocking Predominant in biologics

Particles block a portion of a pore Both particle types

Gradual Pore Blocking

t/V = At + B

A,B,C,D,E & k are constants t = time, V = cumulative volume

Filter Plugging Models - Vmax

Mechanism of filter plugging: d2t/dV2 = k(dt/dV)n

where:
t = filtration time V= cumulative volume at time t k = constant whose dimensions are dependent on n

H.P. Grace, "Structure and Performance of Filter Media,"

AICHE Journal 2(3), 307-336 (1956)

In practical terms:
t/v = t/Vmax + 1/Qi
where: Vmax = maximum volume that can be filtered at time infinity Qi = instantaneous initial flow

Vmax: test procedure

Test Operation:
Select test filter type for application Select test system & scaledown device Wet/vent test filter with buffer or product Gently add representative feed solution to reservoir Filter at constant pressure Record cumulative filtrate weight or volume vs time Stop after either set time, set L/m2 throughput, or set %flow decay
Air Supply

Scaledown filter

Collection Reservoir

Feed Reservoir

Balance

Vmax: data analysis

Vmax analysis

Plot data as t/V vs t Use linear least squares to generate a best fit line If poor fit (r2 < 0.99) or negative slope
Eliminate suspect points (initial wetting or buffer flush out transients, scale jumps,) Re-run, run longer (higher % flow decay) Try fitting another plugging model
min/mL T/V

0.030 CGW1 media 13.8 cm2 0.025 SHF buffer 13.8 cm2 0.020 0.015 0.010 0.005 0.000 0 10 20 30 minutes T 40 50 60

Gradual pore plugging model gives t/V=t/AVmax+1/AQi

If good fit (r2 > 0.99), i.e. CGW1

t/V=0.000209t+0.00979 min/ml Extract model parameters:
Vmax= 1/slope/area= 0.000209 ml-1/13.8 cm2 = 3470 L/m2 Note Vmax -> as slope->0 Qi= 1/intercept/area=1/0.00979 ml/min/13.8 cm2 = 4440 LMH

Application of Vmax
Ideal for rapid screening of membrane and membrane combinations Useful for initial system sizing estimates Three process scenarios or cases are usually relevant: Case 1: Batch Volume to be filtered Case 2: Batch Volume to be filtered at a maximum allowable process time Case 3: Batch Volume to be filtered in a process time with a specified minimum allowable flow rate Largest surface area that fulfills all process requirements is selected Compare area, Amin, to available filter configurations & select smallest configuration that meets/exceeds Amin this is Aconfiguration

Suggested minimum safety factor ~ 20%

Vmax: Sizing Equations

Case 1. Only VB (Batch Volume is given; No batch time, minimum flow)

Amin

VB (Vmax / Atest )

CAUTION!!: DO NOT use with NonPlugging Streams (Vmax >1000).Assume a batch time and use equation under 2 below

Eq. gives the minimum area required; No safety factor is included Ensure that Amin leads to respectable batch times
Case 2. VB (Batch Volume) and tB (Batch time) are given

Amin

VB (Vmax / Atest )

VB (Qi / Atest ) t B

Case 3. VB (Batch Volume), tB (Batch time) & Qmin (minimum flow rate) are given

Qmin (Qi / Atest ) Amin

(Vmax

VB / Atest ) Amin

Using Trial & Error, Solve Eq. For Amin Select the largest Amin from 1., 2., & 3.

Vmax Magnitudes
Vmax Range >1,000 l/m2

Low Plugging Primarily flux based sizing Buffers, simple media Moderate Plugging Vmax critical range Protein solutions, some media High Plugging Cell harvest Serum, hydrolysates

200 l/m2 to 1,000 l/m2

<200 l/m2

Vmax - Summary
Ideal for rapid screening of multiple membrane candidates Minimum of time and feed required Ideal for determining effectiveness of prefilter candidates Good tool for initial filter system sizing Attention to dP, filter area, and feedstream consistency helps minimize scaling error Does not tell you anything about filtrate quality Indirectly Vmax, with a tighter filter, on the filtrate is a measure of filtrate quality Only applies to gradual pore plugging model

Pmax & Tmax: test procedure

Application:
High permeability filters (depth, open prefilters) Filter plugging or filtrate quality limits performance
Pump

Pressure

Scaledown filter Quality measure Collection Reservoir Feed Reservoir Balance

Test Operation:
Select test filter type for application Select test system & scaledown device Wet/vent test filter with buffer or product Gently add representative feed solution to reservoir Filter at constant flow Record volume, pressure drop & filtrate quality (e.g. NTU, 0.2 um Vmax) vs time Stop after test end-point (time, L/m2 throughput, psid pressure drop, filtrate quality)

Centrate Clarification 0.027m2, 110 LMH

30 25 20
psid

50 45 40

15 10 5 0 0 50 100 150 200 minutes time

10 5 0 250

NTU

A1HC psid A1HC NTU

35 30 25 20 15

Scaling up

Rapid scaledown testing

Batch testing Vmax, Pmax, Tmax Filter screening Extrapolate process & rough sizing to time, throughput to variable flows/pressures to series operation Sensitivity and variability testing Include scaling & safety factors

Simulation scaledown testing

Full filtration process Time, throughput Variable flows/pressures Single/series Monitor P for each filter, flow rate, throughput (turbidity for depth filters) Validate batch testing Update process & sizing Flow & capacity Pilot test: area, feed volume

Simulation pilot testing

Full Filtration process Prepare to replace filters that plug Attention to all steps (flush, SIP, integrity) Monitor P for each filter, flow rate, throughput (turbidity for depth filters) Validate scaledown Updated process & sizing Flow & capacity Area

Engineering runs at scale

Vmax References

System Design Considerations

Sterilization and Integrity Testing

Sterilization Options
Autoclave Steam-in-place Gamma

Post sterilization assembly

Likely

None

Can vary from multiple steps to none

Operation

Automated

Can vary from fully manual to fully automated

Extensive

Vendor

Validation

Extensive

Vendor

Filter damage

Extremely remote

Design/operation Materials dependent dependent

Options for Process Operation Sequence

Set up & Install Cartridges

SIP

Flush/Wet

Process

Test Integrity

Flush/Wet

Air Blow Down P < 5 psi

Test Integrity

Operation Sequence Options

SIP > Wet > IT Advantages Identify filters damaged during SIP before operation Constraints Filter must wet easily after SIP Wet > IT > SIP Advantages Perform IT under non-sterile conditions Constraints Must be able to SIP wet filter

Successful SIP Considerations

Steam pressure and differential pressure must be controlled to assure sterility and prevent damage to filter
Filters must be resistant to hydraulic and thermal stress Integrity failures equal:
Lost time 2 to 4 hours for re-installation and re-SIP Potential contamination resolution varies widely from thousands to potentially millions of dollars

Process system SIP is a time-consuming operation

Being able to SIP multiple system components together saves time
Minimizes potential for sterility breaches Save labor time

SIP Options Filter + tank - separate sterilization

Sterilize filter and tank separately or consecutively Sterile boundary Long
V3 V1 P1 Transfer P4 line

Steam
P2

Vent filter

More equipment Complex

Air/N2

P3

Steam
V2
T3 T1

Multi-Round Experiments Forward or Reverse SIP

Dry Forward SIP

Express SHC CHGE73TS3 Multi-Round 3 x 30" Filter Wetting, Integrity Test, Blow Down, and "Dry" Forward SIP at 121 oC, 20 minute Exposure Time
140
Integrity Test Blow Down Spore Installation SIP Exposure

100 90 80
Differential Pressure (psid)

120

Wetting

100
Temperature (oC)

70 60 50

80

60

40 30 20

40

20 10 0 0 15 30 45 60 75 90 105 120 135 150 165 180 195 210 225 240 255 270 285 300 Time (Min) TC- A 01 TC- A 09 TC- A 02 TC- A 10 TC- A 03 Delta P TC- A 04 TC- A 05 TC- A 06 TC- A 07 TC- A 08 0 315

Wet Forward SIP

Express SHR with Prefilter CHVE73TS3 Multi-Round 3 x 30" o Filter Wetting, Integrity Test, and "Wet" Forward SIP at 121 C, 20 minute Exposure Time
140
Wetting Integrity Test Spore Installation SIP Exposure

100 90 80
Differential Pressure (psid)

120

100
Temperature (oC)

70 60 50

80

60

40 30 20

40

20 10 0 0 15 30 45 60 75 Time (Min) TC- A 01 TC- A 09 TC- A 02 TC- A 10 TC- A 03 Delta P TC- A 04 TC- A 05 TC- A 06 TC- A 07 TC- A 08 90 105 120 135 0 150

SIP Options
Filter + tank - Reverse steaming
P3

Easy, maximum simplicity Reduced number of drain & valves Clean steam required

Liquid

filter
P1

P2

Vent filter

Requires a robust filter

T1

Dry Reverse SIP

Express SHC CHGE73TS3 Multi-Round 3 x 30" Filter Wetting, Integrity Test, Blow Down, and "Dry" Reverse SIP at 121 oC, 20 minute Exposure Time
140
Wetting Integrity Test Blow Down Spore Installation SIP Exposure

100 90 80
Differential Pressure (psid)

120

100
Temperature (oC)

70 60 50

80

60

40 30 20

40

20 10 0 0 15 30 45 60 75 90 Time (Min) TC- A 01 TC- A 09 TC- A 02 TC- A 10 TC- A 03 Delta P TC- A 04 TC- A 05 TC- A 06 TC- A 07 TC- A 08 105 120 135 150 165 0 180

Wet Reverse SIP

Express SHC CHGE73TS3 Multi-Round 3 x 30" o Filter Wetting, Integrity Test, and "Wet" Reverse SIP at 121 C, 20 minute Exposure Time
140
Wetting Integrity Test Spore Installation SIP Exposure

100 90 80
Differential Pressure (psid)

120

100
Temperature (oC)

70 60 50

80

60

40 30 20

40

20 10 0 0 15 30 45 60 75 90 Time (Min) TC- A 01 TC- A 09 TC- A 02 TC- A 10 TC- A 03 Delta P TC- A 04 TC- A 05 TC- A 06 TC- A 07 TC- A 08 105 120 135 150 165 0 180

SIP Cycle Summary

Forward, Dry Best option for minimum filter stress Reverse, Dry Best option for simultaneous SIP of filters and tanks Forward, Wet OK if steam introduction is carefully controlled Reverse, Wet Not recommended

Post Sterilization/Pre-use Integrity Testing

Regulatory references
FDA Aseptic Process Guidelines, 2004

EMEA Annex 1, 2008

Practical Challenge Sterile Barrier Options

Process equipment Catch Can Millidisk Barrier Filter

Redundant filter system testing Cost/Benefit Considerations

Regulatory Guidance - FDA

FDA 2004 Aseptic Processing Guidelines

Regulatory Guidance - FDA

Regulatory Guidance - FDA

Regulatory Guidance - EMEA

EMEA Annex 1 2008

Regulatory Guidance - EMEA

Regulatory Guidance - EMEA

Regulatory Guidance Summary

Documents & Guidance provide general, not practical guidance Vendors and PDA Technical Report 26, Sterilizing Filtration of Liquids are good sources of practical guidance
EU / EMEA says less but appears far more skeptical than FDA

White Paper for each company on qualification and validation of aseptic processes is very prudent.

Practical Challenges
Condensate

Steam

Water

Gas
Class C Class B

Remove
Condensate Wetting Liquid Test Gas

Sterilising Product Filter

Maintain downstream
Sterility Atmospheric pressure (test)

Filling Line A

Current Practice
Fluids are sent to downstream equipment
Vented process tank or filling manifold Good option when product wet filter testing is used

Catch can with a sterile vent filter

Not easy to handle Separate autoclaving No filter blow-down One vent filter to integrity test Limited wetting volume

to filling

An Alternate Solution
Millidisk Barrier Filter technology facilitates
Integrity testing of product filters in-situ post sterilization before use Sterile equipment Draining (residual condensate) Cooling Drying Venting (maintain atmospheric pressure on the sterile side)

Without breaching sterility

Millidisk Barrier Filter Design

One disk pair

Liquid Condensate
3 Hydrophilic membranes

1 Hydrophobic membrane

Gas

Using the Millidisk Barrier Filter

1 SIP

Wetting

3 Testing
Integrity test of liquid filter

Atmospheric Pressure

Steam

Water
1 bar max

After SIP, cool down with compressed air Eliminate condensate, steam & air through barrier filter
Open Closed

Once cool, wet the sterilizinggrade filter and direct the effluent to drain through the Barrier Filter.

Vent test gas through Barrier Filter

Using the Millidisk Barrier Filter

Blowdown

5 a Process

Compressed air

Product

After integrity of Barrier Filter is confirmed, start processing.

Dry the liquid filter prior to processing. Vent gas through the Barrier Filter.

5b

Integrity test Barrier Filter off-line, IPA 70/30 bubble point test.

Redundant Sterile Filtration

Aervent
P1 P2

P3

MilliBarrier
T

Post Sterilization/Pre-use IT Options

Advantages Disadvantages

Process Tanks

No additional equipment

Product IT specs needed Entire system requires cleaning and re-prep in case of filter failure Lost product No post-SIP extractables flush Flush volume limited Additional equipment to assemble and sterilize Likely no allowance for re-test Vent filter must be tested Additional equipment to assemble and sterilize Additional filter to test Limited flow rate (limit is 30-inch filters)

Catch-Can

Use water test specs Extractables flush

Barrier Filter

Water test specs Extractables flush No limit to flush volume facilitates re-test if necessary Easy Filter blow-down

Many Examples

Barrier Filter technology currently in use in a wide range of processes

Post SIP/Pre-use Integrity Testing Cost/Benefit Considerations

Costs Additional Capital Costs Hardware Hardware validation Processing costs Product to drain More filters to buy, prep and test More time for filter testing More cost associated with false IT failure Added Risks Sterility compromised by additional manipulations around the sterilized system Frequency unknown although expected to be low in a well designed and operated system Benefits Simple EU regulation compliance Avoid processing with SIP damaged filter Value of lost product Cost of re-work (if allowed) GCC analysis suggests SIP related filter damage occurs about 1/15000 batches
For all processes worldwide Actual for site would be predicted from site specific history

Redundant filtration essentially eliminates this risk assuming one filter is validated for sterilization

Conclusion
Designing successful sterilizing filtration systems requires Knowledge of filter/fluid interactions Retention and plugging mechanisms Sizing techniques and applicability Sterilization methods Selection based on materials and facility Understand limitations for successful SIP implementation Integrity testing Understand risk/benefits of various options
Including regulatory requirements

Vendor experience is critical for efficient implementation

Acknowledgements
Herb Lutz Kerry Roche-Lentine Richard Morin Maurice Phelan